中国癌症杂志 ›› 2015, Vol. 25 ›› Issue (10): 791-795.doi: 10.3969/j.issn.1007-3969.2015.10.006

• 论著 • 上一篇    下一篇

miR-101靶向甲基化转移酶3A抑制人卵巢癌细胞生长与侵袭

胡可可,邓赫男,谭 琛,彭丽秀,肖斌梅   

  1. 郴州市第一人民医院妇产科,湖南 郴州 423000
  • 出版日期:2015-10-30 发布日期:2015-12-17
  • 通信作者: 胡可可 E-mail:176024235@qq.com
  • 基金资助:
    湖南省教育厅课题资助项目(13C929)。

miR-101 inhibits growth and invasion of ovarian cancer cells by targeting DNMT3A

HU Keke, DENG Henan, TAN Chen, PENG Lixiu, XIAO Binmei   

  1. Department of Gynaecology and Obstetrics, First Hospital of Chenzhou, Chenzhou 423000, Hunan, China
  • Published:2015-10-30 Online:2015-12-17
  • Contact: HU Keke E-mail: zrzl67@aliyun.com

摘要: 背景与目的:miR-101在胃癌、结肠癌、乳腺癌以及前列腺癌中表达下调,有类似抑癌基因样作用,然而,其在卵巢癌中的作用尚未明确。该研究旨在探讨miR-101是否通过靶向调控甲基化转移酶3A(DNMT3A)抑制人卵巢癌细胞生长与侵袭,从而进一步揭示miR-101的抑瘤机制。方法:采用实时定量聚合酶链式反应(quantitative real-time polymerase chain reaction,qRT-PCR)检测22例卵巢癌组织及癌旁正常卵巢组织中miR-101的表达改变;将miR-101 mimics转染于卵巢癌SKOV3细胞,以DNMT3A siRNA为阳性对照,采用蛋白[质]印迹法(Western blot)检测外源过表达miR-101对DNMT3A蛋白表达水平的影响;采用噻唑蓝(thiazolyl blue,MTT)和Transwell侵袭实验检测外源高表达miR-101对人卵巢癌细胞生长与侵袭能力的影响。结果:qRT-PCR检测结果显示,miR-101在22例卵巢癌组织中的表达水平较癌旁正常组织明显下调;Western blot检测结果显示,外源过表达miR-101或沉默DNMT3A能下调SKOV3细胞DNMT3A蛋白的表达水平;MTT检测结果显示,转染miR-101 mimics或沉默DNMT3A 48、72和96 h后D值与对照组比较明显减少,差异均有统计学意义(P<0.05);Transwell侵袭实验显示,转染miR-101 mimics或沉默DNMT3A 36 h后穿过基底膜的细胞数分别为(105±7)个和(107±13)个,与对照组(213±11)个比较能明显减缓SKOV3细胞的穿膜能力,差异有统计学意义(P<0.05)。结论:miR-101通过靶向调控DNMT3A抑制人卵巢癌细胞生长与侵袭。

关键词: miR-101, 卵巢癌, 甲基化转移酶3A, 生长和侵袭

Abstract: Background and purpose: miR-101 has been reported to be down-regulated in gastric cancer, colorectal cancer, breast cancer as well as prostate cancer acting as a tumor suppressor gene. However, its function in ovarian cancer is still unknown. The aim of this study was to investigate whether miR-101 can suppress cell growth and invasion of ovarian cancer cells by targeting DNMT3A, so as to reveal molecular mechanism to inhibit ovarian cancer. Methods: Quantitative real-time palymerase chain reaction (qRT-PCR) method was employed to detect the expression of miR-101 in ovarian cancer and cancer adjacent normal ovarian tissues. SKOV3 cells were transfected with miR-101 mimics, and DNMT3A siRNA was transfected as a positive control. Then Western blot was used to detect the expression of DNMT3A protein regulated by miR-101 in SKOV3 cells. The growth and invasion ability of SKOV3 cells were evaluated by MTT and Transwell invasion assays. Results: qRT-PCR showed that miR-101 was down-regulated in ovarian cancer tissues. Western blot showed that the level of DNMT3A protein was inhibited by restored miR-101 or knock-down of DNMT3A in SKOV3 cells. Following transfection of miR-101 mimics or knock-down of DNMT3A for 48, 72 and 96 h respectively, MTT assay showed that the D values were significantly lower than the control group, (P<0.05). After transfection of miR-101 mimics or knock-down of DNMT3A for 36 h, Transwell invasion assay showed that the numbers of cells through the basement membrane was (105±7) and (107±13), respectively, which are significantly different from the control group (213±11), indicating invasion of SKOV3 cells significantly slowed down (P<0.05). Conclusion: miR-101 suppresses cell growth and invasion by targeting DNMT3A in ovarian cancer.

Key words: MiR-101, Ovarian cancer, DNMT3A, Growth and invasion